Manipulation of arterial-venous identity

ABSTRACT

Methods and compositions for manipulating the arterial-venous identity of endothelial cells are provided. The methods comprise introducing an arterial molecular program into endothelial cells of a vein section such that the endothelial cells can remodel to form arterial endothelial cells. The arterial molecular program can comprise one or more polynucleotides encoding various genes that are associated with arterial development and/or differentiation from veins. Expression vectors comprising the genes can be used to introduce the molecular program into the cells. A method of treating a patient having an obstructed blood vessel is also provided.

REFERENCE TO PREVIOUS APPLICATION

[0001] This application claims the benefit of U.S. ProvisionalApplication Serial No. 60/222,759 filed on Aug. 3, 2000.

FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] The U.S. Government may have rights in the present inventionpursuant to the terms of grant number HL65648-01 awarded by the NationalInstitutes of Health and grant number HL03490-01 awarded by the NationalHeart, Lung and Blood Institute.

FIELD OF THE INVENTION

[0003] The present invention relates to methods and compositions formanipulating the arterial-venous identity of endothelial cells. Moreparticularly, the invention relates to methods of inducing arterialmorphology in a vein by transferring an appropriate polynucleotide intoendothelial cells of the vein. Further, the invention relates to methodsof treating a patient having an obstructed blood vessel.

BACKGROUND OF THE INVENTION

[0004] Obstruction of blood vessels diminishes the ability of the vesselto deliver blood to downstream organs, which impacts the long-termhealth of the organ and its host. These obstructive disorders aregenerally referred to as arteriosclerosis. Atherosclerosis, a specificform of arteriosclerosis, primarily affects the aorta and the coronaryarteries.

[0005] Replacement of the obstructed vessel with a graft of some type isthe mainstay of surgical treatment for obstructive vascular disease. Acommon example of this type of procedure is coronary artery bypassgrafting (CABG). This procedure is aimed at alleviating poor bloodperfusion of the heart that results from obstructed coronary arteries,and represents the great majority of vessel replacement procedures inhumans. In a CABG procedure, a surgeon removes an obstructed segment ofthe coronary artery and replaces it with a graft.

[0006] Choice of graft in any vessel replacement procedure is affectedby numerous factors, including rejection by the host immune system,vessel size, and ease of harvesting and handling. For CABG procedures,autologous vessels, particularly the saphenous vein, are predominantlyused as the replacement vessel. The saphenous vein, which ascends alongthe inner side of the leg, is relatively easy to harvest and has asuitable cross-sectional size relative to the coronary arteries.Furthermore, as an autologous vessel, tissue rejection concerns areeliminated. Considering these advantages, it is not surprising thatreplacement of an obstructed coronary artery section with a section ofan autologqus saphenous vein has become a common surgical technique forCABG procedures.

[0007] Despite these advantages, significant drawbacks remain. Forexample, recent estimates indicate that as many as 30% of patients whorequire a CABG procedure do not have veins suitable for grafting. (SeeBourassa, Curr. Opin. Cardiol. 9: 685-691 (1994)). Furthermore,approximately 50% of venous bypass grafts are no longer patent, i.e.,structurally intact, ten years after grafting, (See Edwards, et al.,Surg. Gynecol. & Obstet. 122: 37-42 (1996). The loss of patency of thegraft has serious consequences: at a minimum, it can create a need foran additional surgical procedure and, as a worst case, can lead to heartdamage and death. Considering these limitations and the many benefits ofbypass grafting, there is tremendous interest in improving the patencyof vascular grafts.

[0008] The use of a vein segment in place of an artery segment probablycontributes to the loss of patency in grafts. Arteries have variousstructural features that are not present in veins. For example, whileveins are typically composed of a single layer of endothelium surroundedby a relatively low number of vascular smooth muscle cells, theendothelium of arteries are surrounded by alternating rings of elasticlamellae and vascular smooth muscle cells. These structural differencesallow arteries to accommodate different physiological conditions thanveins. For example, arteries are typically under higher hemodynamicstress (70-105 mm Hg) than veins (0-8 mm Hg).

[0009] Because of these structural features, the use of arteries asgrafts has been explored. Indeed, a relatively high percentage ofinternal mammary artery (IMA) grafts remains intact for years after thegrafting procedure. (See Barner, et al., J. Thorac. Cardiovasc. Surg.90:668-75 (1985)). Thus, the use of arteries appears to have advantagesover the use of vascular grafts. Unfortunately, arteries are frequentlydifficult to harvest. Various technical difficulties are associated withpreparation of the IMA and other arteries, such as the gastroepiploicand splenic arteries, placing these vessels in disfavor as replacementgrafts.

[0010] An ideal graft for replacement of an obstructed section of anartery could be a vessel that combines the benefits of veins, such asease of harvesting, with those of arteries, such as the above-mentionedstructural features.

[0011] During embryonic development, endothelial tubes have the capacityto develop into both veins and arteries. The endothelial tubes acquire aspecific identity as either an artery or vein prior to the developmentof the structural features that distinguish the two types of vessels.Molecular programs, comprising various genes and gene products, regulatethe identity of these vessels as either arterial or vascular tissue.However, the mere replacement of a vein under arterial hemodynamicconditions does not lead to the transformation of the vein into anartery.

SUMMARY OF THE INVENTION

[0012] The present invention provides methods of inducing arterialmorphology in a vein. The method comprises changing the arterial-venousidentity of endothelial cells in a segment of a vein to resemble that ofendothelial cells in arterial tissue. With an arterial identity, thecells and surrounding tissue can undergo endothelial remodeling suchthat the vein develops the morphology of an artery, which can improvethe ability to serve as vessel replacement grafts.

[0013] In a preferred embodiment, the method comprises changing thearterial-venous identity of endothelial cells in vascular tissue bytransferring an appropriate polynucleotide(s) into the endothelialcells. The polynucleotide(s) encodes a gene or genes capable of inducingendothelial remodeling of the cells such that the cells resembleendothelial cells associated with an artery. Preferred genes for use inthis method include those that function to allow arteries and veins todevelop distinct identities, such as endoglin and activin receptor-likekinase 1 (Alk-1), and those that are differentially expressed inarteries and veins, such as ephrin-B2, Eph B4, elastin and CD34. Thesegenes can be used individually or in any combination. By introducing anappropriate polynucleotide, the endothelial cells of the vascular tissuecan remodel and transform their structure to those of an artery.

[0014] Furthermore, the present invention also provides methods oftreating a patient having an obstructed blood vessel, such as a patientpresenting atherosclerosis. In a preferred embodiment, the methodcomprises harvesting a section of a vein, such as a section of anautologous saphenous vein, changing the arterial-vascular identity ofthe section by transferring an appropriate polynucleotide into theendothelial cells of the section, removing the obstructed section of avessel, such as a coronary artery, and grafting the section having thechanged arterial-vascular identity for the obstructed section.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION

[0015] The following description of various preferred embodiments of theinvention provides examples of the present invention. The embodimentsdiscussed herein are merely exemplary in nature, and are not intended tolimit the scope of the invention in any manner. Rather, the descriptionof these preferred embodiments serves to enable a person or ordinaryskill in the relevant art to practice the present invention.

[0016] In one embodiment, the present invention provides methods andcompositions for manipulating the arterial-venous identity ofendothelial cells. Manipulation of the arterial-venous identity isaccomplished by transferring one or more polynucleotides, or productsthereof, that encode one or more genes capable of inducing remodeling ofthe cells such that the cells resemble endothelial cells associated withan artery. The polynucleotide encodes genes that belong to one or bothof the following classes:

[0017] 1) genes that have a function of allowing endothelial cells todevelop distinct artery or vein identities; and

[0018] 2) genes that are differentially expressed in endothelial cellsof arteries and veins.

[0019] The development of arterial-venous identity is one step in thepathway that allows embryonic endothelial tubes to develop into eitherof these types of vessels. Furthermore, the inventor has discovered thatinsertion of the polynucleotide described above into venous endothelialcells allows the cells to remodel into arterial endothelial cells. Thedevelopment of an arterial identity is measured by the appearance ofarterial structural features.

[0020] The first class of genes that can be used comprises those genesthat have a function of allowing endothelial cells, during embryonicdevelopment, to develop distinct identities as being either arterial orvenous. During development, vascular remodeling and endothelialmaturation produce the final vasculature system. To form an organizedvascular network, a hierarchy of major and minor vessels thatefficiently transport blood to and from tissues must be established. Theformation of a mature hierarchical vascular system forms in two steps.The first step involves the differentiation, rapid proliferation andtube formation of endothelial cells. This process results in theformation of a meshwork of interconnected and homogeneously sizedendothelial tubes. In the second stage, vascular remodeling andendothelial maturation occurs. Endothelial tubes must be distinguishedas arterial or venous, and an organized network is formed throughdifferential growth, apoptosis, and sprouting of endothelial tubes. Thisprocess of remodeling leads to a well-defined vascular network thatefficiently supplies blood to and removes waste from the target tissueor tumor. Endoglin and activin receptor-like kinase 1 (Alk1) function toinitiate the switch from stage 1-endothelial differentiation and rapidproliferation and stage 2-endothelial maturation and vascularremodeling. (See generally, Ferrara N, and Alitalo, K. Nat. Med.5:1359-64 (1999), Folkman J., and D'Amore P. A., Cell 87:1153-1155(1996), Gale N. W., and Yancopoulos G. D., Genes Dev 13(9):1055-66(1999), Inducing Alk-1 and endoglin function will promote maturation andthe molecular program needed to distinguish arteries from veins.Disrupting the function of these genes will prevent a nascentendothelial network from forming a highly organized network. Thus,blocking the function of these genes may prevent pathologicneovascularization, processes that are critical for cancer growth anddiabetic retinopathy. See, for examples, Li, D. Y., et al., Science284:1534-1537 (1999) and Urness, L. D., et al., Nature Genetics26:328-331 (2000). Examples of genes belonging to this first classinclude endoglin and activin receptor-like kinase 1 (Alk-1).

[0021] Endoglin is a transforming growth factor-β (TGF-β) bindingprotein expressed on the surface of endothelial cells. TGF-β signalingis required for vasculogenesis, the first stage of vascular development.During vasculogenesis, the primary capillary network, composed ofinterconnected and homogenously sized endothelial tubes, is formed.Indeed, mice lacking endoglin die at an early age due to defectivevascular development characterized by poor smooth muscle development andarrested endothelial remodeling. Consequently, endoglin is essential forthe second stage of vascular development, angiogenesis, in which theprimary endothelial network is remodeled into a mature circulatorysystem. See generally Li, D. Y., et al., Science 284:1534-1537 (1999).

[0022] The cDNA encoding endoglin has been described. (Gougos, A. andLetarte, M., J. Biol. Chem. 265(15): 8361-8364 (1990)). The sequence isappended hereto as SEQ ID NO. 1.

[0023] The Alk-1 gene encodes a serine/threonine kinase receptor for theTGF-β superfamily of growth factors (ten, Dijke, P., et al, Science 264(5155): 101-4 (1994); ten, Dijke, P., et al., Oncogene 10: 2879-87(1993); Attisano, L. & Wrana, J. L., Cytokines and Growth Factor Reviews7(4): 327-339 (1996)). The receptor encoded by Alk-1 is highly expressedin the endothelium (Roelen, B. A., et al, Dev. Dyn. 209(4): 418-30(1997)). Also, loss-of-function mutations of Alk-1 are responsible for ahuman vascular dysplasia characterized by arteriovenous malformations(Guttmacher, A. E., et al., N. Engl. J. Med. 333(14): 918-924 (1995);Johnson, D. W., et al., Nat. Genet. 13(2): 189-95 (1996)). Furthermore,anatomical, molecular, and functional distinctions between arteries andveins are lost in mice lacking Alk-1. Lastly, Alk-1 is required forsuccessful embryonic development of distinct arterial and venousvascular beds (Id.).

[0024] The cDNA encoding Alk-1 has been described (ten Dijke, P. P., etal., Oncogene 8(10): 2879-2887 (1993)). The sequence appended hereto asSEQ ID NO. 2

[0025] The second class of genes that can be used in the arterialmolecular program comprises those genes that are differentiallyexpressed in the endothelial cells of arteries and veins. As usedherein, the term “differentially expressed” refers to the relativeextent of expression of a gene in an endothelial cell in an artery ascompared to an endothelial cell in a vein. Examples of genes belongingto this second class include ephrin-B2, EphB4, elastin, and CD34. See,for example, Urness, L. D., et al., Nature Genetics 26:328-331 (2000).

[0026] The epprin-B2 gene encodes an arterial specific molecular markerthat is expressed prior to the appearance of any structural orfunctional differences between arteries and veins (Adams, R. H., et al.Gened Dev. 13:3 295-306 (1999), Wang, H. U., et al., Cell 93(5): 741-53(1998)). Also, while mice lacking the ephrin-B2 gene or the gene for theephrin-B2 receptor, EphB4, develop distinct arterial and venous domains,these mice experience defective endothelial remodeling (Id.; Gerety, S.S., et al., Mol Cell 4:403-14 (1999)). Thus, while ephrin-B2 and EphB4are important arterial markers, they do not regulate the specificationof endothelial tubes to become arteries and veins. Indeed, mice lackingthe Alk-1 gene fail to express normal levels of these markers despitethe presence of an extensive endothelial network. (Urness, L. D., etal., Nature Genetics 26: 328-331 (2000)).

[0027] The cDNA encoding ephrin-B2 has been described. (Bennett, B. D.,et al., Proc. Natl. Acad. Sci. U.S.A. 92(6): 1866-1870 (1995)). Thesequence is appended hereto as SEQ ID NO. 3

[0028] The cDNA encoding EphB4 appended hereto as SEQ ID NO. 4

[0029] Elastin is the main component of the extracellular matrix ofarteries. Elastin has both structural and developmental roles. Duringarterial development, elastin controls proliferation of smooth muscleand stabilizes arterial structure. Indeed, mice lacking elastin die ofan obstructive arterial disease resulting from subendothelial cellproliferation and reorganization of smooth muscle. (See Li, D. Y., etal., Nature 393:276-280 (1998)).

[0030] The cDNA for elastin has been described (Faszio, M. J., et al.,J. Invest. Dermatol. 91(5) 458-464 (1998). The sequence is appendedhereto as SEQ ID NO. 5.

[0031] The CD34 gene encodes a cell surface glycoprotein that isexpressed in early blood vessels, as well as on various hematopoieticcells (See, Wood, H. B., et al., Blood 90(6): 2300-2311 (1977)).

[0032] The cDNA encoding CD34 has been described (NCBI AnnotationProject, Direct Submission, 7-16-2001). The sequence is appended heretoas SEQ ID NO. 6

[0033] Preferably, transferring the polynucleotide(s) into anendothelial cell having a venous identity is accomplished bytransferring an expression vector comprising one or more of the genesdescribed above. Suitable expression vectors useful in accordance withthe present invention include eukaryotic, plasmid and viral vectors, andcombinations thereof. Examples of useful viral vectors includerecombinant viral vectors such as adenoviral, retroviral, herpesviral,pox viral, and adeno-associated viral vectors. Preferably, thepolynucleotides are contained within the expression vector. Alsopreferable, the expression vector is adapted to introduce thepolynucleotide into the endothelial cells.

[0034] The transferring of the polynucleotide into the endothelial cellscan occur in vivo or ex vivo. Preferably, the transferring occurs exvivo on a vessel segment harvested from a patient. Conventionaltransduction techniques can be utilized to carry out the ex vivotransferring of the polynucleotide into the endothelial cells when viralvectors are used. Examples of suitable transduction techniques includethose described in Kibbe, M. R., et al., J. Vasc. Surg.34(1): 156-65(2001) and Moawad, J., et al., Ann. Vasc. Surg. 15(3):367-73 (2001). Thetransduction should be carried out using a sufficient number of vectorparticles to ensure adequate transferring of the polynucleotide. Also,the transduction should be carried out under culturing conditions thatare conducive to the viability of the endothelial cells as well as thetransduction by the vector. Preferred number of vector particles andlength of transduction period for changing the arterial venous identityof a segment of a saphenous vein are between approximately 1×10⁸ and1×10¹² viral particles for 15 to 45 minutes. Particularly preferable,approximately 1×10¹⁰ to 1×10¹² viral particles are exposed to the veinsegment for approximately 30 minutes. Most preferable, approximately1×10¹¹ viral particles are exposed to the endothelial cells of the veinsegment for approximately 30 minutes.

[0035] The genes may be encoded on a plasmid or other similar constructand then incorporated into the vector. Conventional molecular biologytechniques can be employed to create suitable constructs for use in thepresent invention.

[0036] Preferred viral vectors include recombinant retroviral andadeno-associated viral vectors. Recombinant retroviral vectors arefrequently used for gene transfer, and methods for constructing suchvectors are known in the art (Hodgson, Bio/Technology 13: 222-225(1995); Miyanohara, et al., Proc. Natl. Acad. Sci. USA 85: 6538-6542(1988); Rosenberg, et al., New Engl. J. Med. 323: 570-578 (1990)).Preferably, retroviral vectors with impaired ability to replicate andtransform are used.

[0037] Methods for producing recombinant adeno-associated viral (AAV)vectors are also known in the art. Briefly, a suitable producer cellline is transfected with an AAV vector containing the gene of interest,which can be encoded on a plasmid. AAV helper functions (i.e., theproducts of the AAV rep and cap genes) and accessory functions, whichare typically derived from a helper virus, such as adenovirus orherpesvirus, are then expressed in the producer cell. Once these factorscome together, the gene(s) of interest is (are) replicated and packagedas though it were a wild-type AAV genome, forming a recombinant virion.When cells, such as endothelial cells, are infected with the resultingAAV virions, the gene(s) of interest enter the cell and is (are)expressed. Because the cells lack the rep and cap genes and the helpervirus accessory function genes, the rAAV are replication defective; thatis, they cannot further replicate and package their genomes. Similarly,without a source of rep and cap genes, wild-type AAV cannot be formed inthe infected cell. For a detailed discussion on the production of rAAVvirions, see U.S. Pat. No. 6,001,650 to Colosi for HIGH-EFFICIENCYWILD-TYPE —FREE AAV HELPER FUNCTIONS.

[0038] The polynucleotides encoding the gene(s) of interest can beinserted into the expression vectors and used for cell transfectionusing conventional recombinant techniques, such as those described bySambrook, Fritsch & Maniatis, in “Molecular Cloning, A LaboratoryManual” (2d ed): pp. E.5 (Cold Spring Harbor Press, Cold Spring Harbor,N.Y. 1989). Alternatively, the expression vectors can be prepared usinghomologous recombination techniques, such as those described byDavidson, et al., Nature Gen. 3: 219-223. (1993) and Lemarchand, Proc.Natl. Acad. Sci. USA 89(14): 6482-6486 (1992).

[0039] The expression vectors of the present invention can additionallycontain regulatory elements such as promoters, as well as selectionmarkers, such as antibiotic resistance genes. Furthermore, theexpression vectors can include tags that allow for binding of theprotein of interest to a binding agent of some sort, which can be usedto facilitate purification and/or localization and targeting efforts.Various such tags are known to those skilled in the art. Examplesinclude F_(c) receptors and Hexo-histidine tags.

[0040] It is well established that viral vectors will be taken up intoand integrated into cells in vivo, to eventually express the viral DNA,including any inserted constructs (Nabel, U.S. Pat. No. 5,328,470;Yoshimura, et al., J. Biol. Chem. 268(4): 2300-2303 (1993); Crystal, AM.J. Med. 92(6A): 445-525 (1992); Lemarchand, et al., Proc. Natl. Acad.Sci. USA 89(14): 6482-6486 (1992)).

[0041] Alternatively, non-viral methods can be used to introduce thepolynucleotides into the endothelial cells. Essentially, any suitablemethod for introducing DNA into cells for later expression can beutilized. For example, techniques such as calcium phosphateco-precipitation (Graham, et al., Virol. 52: 456-467 (1973)), directmicro-injection of DNA into cells (Capecchi, Cell 22: 479-488 (1980)),lipisome-mediated gene transfer (Mannino, et al., BioTechniques 6:682-690 (1988), lipid-mediated transfection (Felgner, et al., Proc.Natl. Acad. Sci. USA 84: 7413-7417 (1987)), delivery using DNA-coatedstents placed at a target site by a catheter, and nucleic acid deliveryusing high-velocity microprojectiles (Klein, et al., Nature 327: 70-73(1987)) can be used. Furthermore, electroporation methods forintroducing DNA into cells and tissues can be used (Shigekawa, et al.,BioTechniques 6: 742-751 (1988)).

[0042] Those skilled in the art will readily recognize that the variousgenes introduced into endothelial cells, using either viral or non-viralmethods, may be operably linked to control elements such as promotersand enhancers, that are capable of driving or repressing gene expressionunder appropriate conditions. Termination signals, such aspolyadenylation sites, can also be included. Control elements, such asinducible promoters, that allow controlled expression of the gene ofinterest are available. For example, an ecdysone-inducible promoter canbe utilized to regulate gene expression. (See, e.g., Stratagenes;Complete Control™ Inducible Mammalian Expression System InstructionManual—available online athttp://www.stratagene.com/manuals/index.shtm). Other examples ofsuitable inducible promoters that are functional in mammalian cellsinclude those that are induced (or repressed) by tetracycline and itsderivatives, RU486, and rapamycin and its derivatives (See, e.g. Grossen& Brujard, Proc. Natl. Acad. Sci, USA 89: 5547-5551 (1992); Wang, etal., Gene Therapy, 4: 432-441 (1997); and Riviera, et al., NatureMedicine 2: 1028-1032 (1996).

[0043] The manipulation of the arterial-vascular identity of endothelialcells may also be accomplished by introducing the products of one ormore of the above-mentioned genes into the endothelial cells, The geneproducts may be produced using standard recombinant techniques known tothose skilled in the art (See, generally, Sambrook, et al., (supra.)).Recombinantly produced gene products may be purified using conventionalpurification schemes, such as affinity chromatography, size-exclusion,filtration, precipitation, and other suitable techniques. The geneproducts can be introduced into the endothelial cells using techniquessuch as microinjection and protein transduction (see, e.g., Schwarze, etal, Science 285: 1569-1572 (1999)).

[0044] The present invention also provides a composition comprising ablood vessel, such as a section of a vein or a vascular graft, havingendothelial cells comprising an exogenously supplied polynucleotideencoding a gene that is capable of inducing endothelial remodeling. Thegene can be any of the genes described above in the description of themethods of the present invention. Thus, the gene can comprise endoglin,Alk-1, ephrin-B2, EphB4, elastin, and/or CD34. The compositions of thepresent invention are useful as grafts to replace sections of arteriescontaining obstructions, such as coronary arteries affected byatherosclerosis.

[0045] In one embodiment, the composition of the present inventioncomprises a section of an autologous vein, i.e., a section of a vein ofthe patient ultimately receiving the graft. The use of autologous tissueeliminates any tissue rejection concerns.

[0046] Any suitable vein can be utilized for the vein section. Thechoice of vein will depend on various factors, such as ease ofharvesting, ability of the vein to tolerate removal of a section, andthe relative capacity of the vessel as compared to that of theobstructed vessel. Preferred veins include the saphenous vein of theleg. Particularly preferable, the vein section comprises a section ofthe internal or long saphenous vein. Sections of these preferred veinscan be readily harvested by surgical techniques known to those skilledin the art.

[0047] Importantly, the vein section must include the endothelial celllayer (endothelium) such that the polynucleotide and gene can beintroduced into the endothelial cells. The polynucleotide can encode anycombination of the genes discussed above. Also, the polynucleotide canbe introduced into the endothelial cells of the vein section accordingto any suitable technique, such as those described above.

[0048] Alternatively, the composition can comprise an engineered bloodvessel comprising endothelial cells.

[0049] Engineered blood vessels are vessels fabricated from tissueengineering procedures. This class of vessel includes synthetic materialin combination with natural cells, such as endothelial cells, as well ascultured vessels produced from natural materials, such as smooth muscleand endothelial cells. Examples of such vessels, as well as techniquesfor their production, can be found in Huynh, T., et al., NatureBiotechnology, 17: 1083-1086 (1999); Niklason, L. E., et al., Science284: 489-493 (1999); L'Heureux, et al, FASEB J. 12: 47-56 (1998); andCampbell, J. H., et al., Cir. Res. 85: 1173-1178 (1999).

[0050] The expression vectors of the present invention can be introducedinto endothelial cells of an engineered blood vessel in the same manneras that described above for segments of natural veins.

[0051] The present invention also provides a method of treating apatient having an obstructed blood vessel. The method of treatment canbe practiced on any mammal, but is particularly well-suited for treatinghumans. The method is particularly well-suited for treating patientshaving obstructed arteries, such as coronary arteries affected byatherosclerosis.

[0052] As detailed above, surgical grafting of a vascular graft in placeof the obstructed artery is a common surgical technique for treatingpatients with obstructed vessels. The method of the present inventioncan be practiced in accordance with guidelines known in the art, such asthose relating to the need for bypass grafting as a function of thefraction of the vessel blocked.

[0053] In a preferred embodiment, the method of treatment comprisesproviding a graft comprising endothelial cells, changing thearterial-vascular identity of the endothelial cells by transferring apolynucleotide encoding a gene capable of inducing endothelialremodeling into the endothelial cells, removing an obstructed section ofa vessel of the patient, and grafting the graft having the endothelialcells with changed arterial-vascular identity in place of the removedobstructed section. The arterial-vascular identity of the endothelialcells can be changed ex vivo prior to grafting, or in vivo aftergrafting.

[0054] The graft can comprise a vein section or an engineered bloodvessel, as described above. If the graft comprises a vein section, thesection preferably comprises a section of an autologous vein, andparticularly preferably comprises a section of a saphenous vein of thepatient. Also, if the graft comprises a vein section, the method mayfurther comprise harvesting the vein section from a vein of the patient.The harvesting can be accomplished according to conventional techniquesknown in the art.

[0055] The changing the arterial-vascular identity of the endothelialcells by transferring an appropriate polynucleotide into the endothelialcells can be accomplished according to the methods of the presentinvention, detailed above.

[0056] The removing an obstructed section of a vessel of a patient andthe grafting of the graft in place of the removed obstructed section canboth be accomplished according to conventional techniques known in theart.

EXAMPLE

[0057] The present invention can be carried out to alter thearterial-venous identity of endothelial cells in a vein segment in an exvivo environment. This preferred method is particularly well-suited fortreating a segment of a vein that has been harvested from a patientsuffering from an obstructed blood vessel. The treated vein segment canbe used as a graft to replace an obstructed section of the obstructedvessel.

[0058] When practicing this method, a segment of a saphenous vein of thepatient will be harvested according to conventional surgical procedures.The section will be dissected to provide a segment that is of a suitablelength, i.e. a length sufficient to allow the segment to serve as areplacement for the obstructed section of the obstructed vessel.

[0059] The vein segment will be passively transduced with an adenoviralvector carrying one or more genes encoding Alk-1, endoglin, ephrin-B2,Eph-B4, elastin, and CD34. The transduction will be carried out usingapproximately 1×10¹¹ adenoviral vector particles for 30 minutes usingstandard techniques. Examples of suitable transduction techniques aredescribed in Kibbe, M. R., et al., J. Vasc. Surg.34(1): 156-65 (2001)and Moawad, J., et al., Ann. Vasc. Surg. 15(3):367-73 (2001).

[0060] The obstructed section of the obstructed vessel will be removedusing conventional surgical techniques. Lastly, the transduced veinsegment will be interposed as a graft in place of the obstructedsection. Thus, if a coronary artery was the obstructed vessel, thetransduced vein segment will be grafted into the coronary circulation inplace of the obstructed section. The grafting can occur in theperipheral circulation, if needed, based on the location of theobstructed section of the obstructed vessel.

[0061] All references cited and otherwise referred to herein are herebyincorporated in their entirety, except to the extent to which they maycontradict any definition or statement herein.

[0062] The foregoing disclosure is the best mode devised by the inventorfor practicing the invention. It is apparent, however, that severalvariations in accordance with the present invention may be conceivableto one of ordinary skill in the relevant art. Inasmuch as the foregoingdisclosure is intended to enable such person to practice the instantinvention, it should not be construed to be limited thereby, but shouldbe construed to include such aforementioned variations. As such, thepresent invention should be limited only by the spirit and scope of thefollowing claims.

1 6 1 3142 DNA Homo sapiens 1 cctgggccgg ccgggctgga tgagccgggagctccctgct gccggtcata ccacagcctt 60 catctgcgcc ctggggccag gactgctgctgtcactgcca tccattggag cccagcaccc 120 cctccccgcc catccttcgg acagcaactccagcccagcc ccgcgtccct gtgtccactt 180 ctcctgaccc ctcggccgcc accccagaaggctggagcag ggacgccgtc gctccggccg 240 cctgctcccc tcgggtcccc gtgcgagcccacgccggccc cggtgcccgc ccgcagccct 300 gccactggac acaggataag gcccagcgcacaggccccca cgtggacagc atggaccgcg 360 gcacgctccc tctggctgtt gccctgctgctggccagctg cagcctcagc cccacaagtc 420 ttgcagaaac agtccattgt gaccttcagcctgtgggccc cgagaggggc gaggtgacat 480 ataccactag ccaggtctcg aagggctgcgtggctcaggc ccccaatgcc atccttgaag 540 tccatgtcct cttcctggag ttcccaacgggcccgtcaca gctggagctg actctccagg 600 catccaagca aaatggcacc tggccccgagaggtgcttct ggtcctcagt gtaaacagca 660 gtgtcttcct gcatctccag gccctgggaatcccactgca cttggcctac aattccagcc 720 tggtcacctt ccaagagccc ccgggggtcaacaccacaga gctgccatcc ttccccaaga 780 cccagatcct tgagtgggca gctgagaggggccccatcac ctctgctgct gagctgaatg 840 acccccagag catcctcctc cgactgggccaagcccaggg gtcactgtcc ttctgcatgc 900 tggaagccag ccaggacatg ggccgcacgctcgagtggcg gccgcgtact ccagccttgg 960 tccggggctg ccacttggaa ggcgtggccggccacaagga ggcgcacatc ctgagggtcc 1020 tgccgggcca ctcggccggg ccccggacggtgacggtgaa ggtggaactg agctgcgcac 1080 ccggggatct cgatgccgtc ctcatcctgcagggtccccc ctacgtgtcc tggctcatcg 1140 acgccaacca caacatgcag atctggaccactggagaata ctccttcaag atctttccag 1200 agaaaaacat tcgtggcttc aagctcccagacacacctca aggcctcctg ggggaggccc 1260 ggatgctcaa tgccagcatt gtggcatccttcgtggagct accgctggcc agcattgtct 1320 cacttcatgc ctccagctgc ggtggtaggctgcagacctc acccgcaccg atccagacca 1380 ctcctcccaa ggacacttgt agcccggagctgctcatgtc cttgatccag acaaagtgtg 1440 ccgacgacgc catgaccctg gtactaaagaaagagcttgt tgcgcatttg aagtgcacca 1500 tcacgggcct gaccttctgg gaccccagctgtgaggcaga ggacaggggt gacaagtttg 1560 tcttgcgcag tgcttactcc agctgtggcatgcaggtgtc agcaagtatg atcagcaatg 1620 aggcggtggt caatatcctg tcgagctcatcaccacagcg gaaaaaggtg cactgcctca 1680 acatggacag cctctctttc cagctgggcctctacctcag cccacacttc ctccaggcct 1740 ccaacaccat cgagccgggg cagcagagctttgtgcaggt cagagtgtcc ccatccgtct 1800 ccgagttcct gctccagtta gacagctgccacctggactt ggggcctgag ggaggcaccg 1860 tggaactcat ccagggccgg gcggccaagggcaactgtgt gagcctgctg tccccaagcc 1920 ccgagggtga cccgcgcttc agcttcctcctccacttcta cacagtaccc atacccaaaa 1980 ccggcaccct cagctgcacg gtagccctgcgtcccaagac cgggtctcaa gaccaggaag 2040 tccataggac tgtcttcatg cgcttgaacatcatcagccc tgacctgtct ggttgcacaa 2100 gcaaaggcct cgtcctgccc gccgtgctgggcatcacctt tggtgccttc ctcatcgggg 2160 ccctgctcac tgctgcactc tggtacatctactcgcacac gcgtgagtac cccaggcccc 2220 cacagtgagc atgccgggcc cctccatccacccgggggag cccagtgaag cctctgaggg 2280 attgaggggc cctggcagga ccctgacctccgcccctgcc cccgctcccg ctcccaggtt 2340 cccccagcaa gcgggagccc gtggtggcggtggctgcccc ggcctcctcg gagagcagca 2400 gcaccaacca cagcatcggg agcacccagagcaccccctg ctccaccagc agcatggcat 2460 agccccggcc ccccgcgctc gcccagcaggagagactgag cagccgccag ctgggagcac 2520 tggtgtgaac tcaccctggg agccagtcctccactcgacc cagaatggag cctgctctcc 2580 gcgcctaccc ttcccgcctc cctctcagaggcctgctgcc agtgcagcca ctggcttgga 2640 acaccttggg gtccctccac cccacagaaccttcaaccca gtgggtctgg gatatggctg 2700 cccaggagac agaccacttg ccacgctgttgtaaaaaccc aagtccctgt catttgaacc 2760 tggatccagc actggtgaac tgagctgggcaggaagggag aacttgaaac agattcaggc 2820 cagcccagcc aggccaacag cacctccccgctgggaagag aagagggccc agcccagagc 2880 cacctggatc tatccctgcg gcctccacacctgaacttgc ctaactaact ggcaggggag 2940 acaggagcct agcggagccc agcctgggagcccagagggt ggcaagaaca gtgggcgttg 3000 ggagcctagc tcctgccaca tggagccccctctgccggtc gggcagccag cagaggggga 3060 gtagccaagc tgcttgtcct gggcctgcccctgtgtattc accaccaata aatcagacca 3120 tgaaacctga aaaaaaaaaa aa 3142 21970 DNA Homo sapiens 2 aggaaacggt ttattaggag ggagtggtgg agctgggccaggcaggaaga cgctggaata 60 agaaacattt ttgctccagc ccccatccca gtcccgggaggctgccgcgc cagctgcgcc 120 gagcgagccc ctccccggct ccagcccggt ccggggccgcgccggacccc agcccgccgt 180 ccagcgctgg cggtgcaact gcggccgcgc ggtggaggggaggtggcccc ggtccgccga 240 aggctagcgc cccgccaccc gcagagcggg cccagagggaccatgacctt gggctccccc 300 aggaaaggcc ttctgatgct gctgatggcc ttggtgacccagggagaccc tgtgaagccg 360 tctcggggcc cgctggtgac ctgcacgtgt gagagcccacattgcaaggg gcctacctgc 420 cggggggcct ggtgcacagt agtgctggtg cgggaggaggggaggcaccc ccaggaacat 480 cggggctgcg ggaacttgca cagggagctc tgcagggggcgccccaccga gttcgtcaac 540 cactactgct gcgacagcca cctctgcaac cacaacgtgtccctggtgct ggaggccacc 600 caacctcctt cggagcagcc gggaacagat ggccagctggccctgatcct gggccccgtg 660 ctggccttgc tggccctggt ggccctgggt gtcctgggcctgtggcatgt ccgacggagg 720 caggagaagc agcgtggcct gcacagcgag ctgggagagtccagtctcat cctgaaagca 780 tctgagcagg gcgacacgat gttgggggac ctcctggacagtgactgcac cacagggagt 840 ggctcagggc tccccttcct ggtgcagagg acagtggcacggcaggttgc cttggtggag 900 tgtgtgggaa aaggccgcta tggcgaagtg tggcggggcttgtggcacgg tgagagtgtg 960 gccgtcaaga tcttctcctc gagggatgaa cagtcctggttccgggagac tgagatctat 1020 aacacagtat tgctcagaca cgacaacatc ctaggcttcatcgcctcaga catgacctcc 1080 cgcaactcga gcacgcagct gtggctcatc acgcactaccacgagcacgg ctccctctac 1140 gactttctgc agagacagac gctggagccc catctggctctgaggctagc tgtgtccgcg 1200 gcatgcggcc tggcgcacct gcacgtggag atcttcggtacacagggcaa accagccatt 1260 gcccaccgcg acttcaagag ccgcaatgtg ctggtcaagagcaacctgca gtgttgcatc 1320 gccgacctgg gcctggctgt gatgcactca cagggcagcgattacctgga catcggcaac 1380 aacccgagag tgggcaccaa gcggtacatg gcacccgaggtgctggacga gcagatccgc 1440 acggactgct ttgagtccta caagtggact gacatctgggcctttggcct ggtgctgtgg 1500 gagattgccc gccggaccat cgtgaatggc atcgtggaggactatagacc acccttctat 1560 gatgtggtgc ccaatgaccc cagctttgag gacatgaagaaggtggtgtg tgtggatcag 1620 cagaccccca ccatccctaa ccggctggct gcagacccggtcctctcagg cctagctcag 1680 atgatgcggg agtgctggta cccaaacccc tctgcccgactcaccgcgct gcggatcaag 1740 aagacactac aaaaaattag caacagtcca gagaagcctaaagtgattca atagcccagg 1800 agcacctgat tcctttctgc ctgcaggggg ctgggggggtggggggcagt ggatggtgcc 1860 ctatctgggt agaggtagtg tgagtgtggt gtgtgctggggatgggcagc tgcgcctgcc 1920 tgctcggccc ccagcccacc cagccaaaaa tacagctgggctgaaacctg 1970 3 2902 DNA Homo sapiens 3 cacagccatg gctgtgagaagggactccgt gtggaagtac tgctggggtg ttttgatggt 60 tttatgcaga actgcgatttccaaatcgat agttttagag cctatctatt ggaattcctc 120 gaactccaaa tttctacctggacaaggact ggtactatac ccacagatag gagacaaatt 180 ggatattatt tgccccaaagtggactctaa aactgttggc cagtatgaat attataaagt 240 ttatatggtt gataaagaccaagcagacag atgcactatt aagaaggaaa atacccctct 300 cctcaactgt gccaaaccagaccaagatat caaattcacc atcaagtttc aagaattcag 360 ccctaacctc tggggtctagaatttcagaa gaacaaagat tattacatta tatctacatc 420 aaatgggtct ttggagggcctggataacca ggagggaggg gtgtgccaga caagagccat 480 gaagatcctc atgaaagttggacaagatgc aagttctgct ggatcaacca ggaataaaga 540 tccaacaaga cgtccagaactagaagctgg tacaaatgga agaagttcga caacaagtcc 600 ctttgtaaaa ccaaatccaggttctagcac agacggcaac agcgccggac attcggggaa 660 caacatcctc ggttccgaagtggccttatt tgcagggatt gcttcaggat gcatcatctt 720 catcgtcatc atcatcacgctggtggtcct cttgctgaag taccggagga gacacaggaa 780 gcactcgccg cagcacacgaccacgctgtc gctcagcaca ctggccacac ccaagcgcag 840 cggcaacaac aacggctcagagcccagtga cattatcatc ccgctaagga ctgcggacag 900 cgtcttctgc cctcactacgagaaggtcag cggggactac gggcacccgg tgtacatcgt 960 ccaggagatg cccccgcagagcccggcgaa catttactac aaggtctgag agggaccctg 1020 gtggtacctg tgctttcccagaggacacct aatgtcccga tgcctccctt gagggtttga 1080 gagcccgcgt gctggagaattgactgaagc acagcaccgg gggagaggga cactcctcct 1140 cggaagagcc cgtcgcgctggacagcttac ctagtcttgt agcattcggc cttggtgaac 1200 acacacgctc cctggaagctggaagactgt gcagaagacg cccattcgga ctgctgtgcc 1260 gcgtcccacg tctcctcctcgaagccatgt gctgcggtca ctcaggcctc tgcagaagcc 1320 aagggaagac agtggtttgtggacgagagg gctgtgagca tcctggcagg tgccccagga 1380 tgccacgcct ggaagggccggcttctgcct ggggtgcatt tcccccgcag tgcataccgg 1440 acttgtcaca cggacctcgggctagttaag gtgtgcaaag atctctagag tttagtcctt 1500 actgtctcac tcgttctgttacccagggct ctgcagcacc tcacctgaga cctccactcc 1560 acatctgcat cactcatggaacactcatgt ctggagtccc ctcctccagc cgctggcaac 1620 aacagcttca gtccatgggtaatccgttca tagaaattgt gtttgctaac aaggtgccct 1680 ttagccagat gctaggctgtctgcgaagaa ggctaggagt tcatagaagg gagtggggct 1740 ggggaaaggg ctggctgcaattgcagctca ctgctgctgc ctctgaaaca gaaagttgga 1800 aaggaaaaaa gaaaaaagcaattaggtagc acagcacttt ggttttgctg agatcgaaga 1860 ggccagtagg agacacgacagcacacacag tggattccag tgcatgggga ggcactcgct 1920 gttatcaaat agcgatgtgcaggaagaaaa gcccctcttc attccgggga acaaagacgg 1980 gtattgttgg gaaaggaacaggcttggagg gaagggagaa agtaggccgc tgatgatata 2040 ttcgggcagg actgttgtggtactggcaat aagatacaca gctccgagct gtaggagagt 2100 cggtctgctt tggatgattttttaagcaga ctcagctgct atacttatca cattttatta 2160 aacacaggga aagcatttaggagaatagca gagagccaaa tctgacctaa aagttgaaaa 2220 gccaaaggtc aaacaggctgtaattccatc atcatcgttg ttattaaaga atccttatct 2280 ataaaaggta ggtcagatccccctcccccc aggttcctcc ttcccctccc gattgagcct 2340 tacgacactt tggtttatgcggtgctgtcc gggtgccagg gctgcagggt cggtactgat 2400 ggagcctgca gcgcccggtgctctgtgtca aggtgaagca catacggcag acctcttaga 2460 gtccttaaga cggaagtaaattatgatgtc cagggggaga aggaagatag gacgtattta 2520 taataggtat atagaacacaagggatataa aatgaaagat ttttactaat atatatttta 2580 aggttgcaca cagtacacaccagaagatgt gaaattcatt tgtggcaatt aagtggtccc 2640 aatgctcagc gcttaaaaaaacaaattgga cagctacttc tgggaaaaac aacatcattc 2700 caaaaagaac aataatgagagcaaatgcaa aaataaccaa gtcctccgaa ggcatctcac 2760 ggaaccgtag actaggaagtacgagcccca cagagcagga agccgatgtg actgcatcat 2820 atatttaaca atgacaagatgttccggcgt ttatttctgc gttgggtttt cccttgcctt 2880 atgggctgaa gtgttctctaga 2902 4 3945 DNA Homo sapiens 4 cgtccacccg cccagggaga gtcagacctgggggggcgag ggccccccaa actcagttcg 60 gatcctaccc gagtgaggcg gcgccatggagctccgggtg ctgctctgct gggcttcgtt 120 ggccgcagct ttggaagaga ccctgctgaacacaaaattg gaaactgctg atctgaagtg 180 ggtgacattc cctcaggtgg acgggcagtgggaggaactg agcggcctgg atgaggaaca 240 gcacagcgtg cgcacctacg aagtgtgtgacgtgcagcgt gccccgggcc aggcccactg 300 gcttcgcaca ggttgggtcc cacggcggggcgccgtccac gtgtacgcca cgctgcgctt 360 caccatgctc gagtgcctgt ccctgcctcgggctgggcgc tcctgcaagg agaccttcac 420 cgtcttctac tatgagagcg atgcggacacggccacggcc ctcacgccag cctggatgga 480 gaacccctac atcaaggtgg acacggtggccgcggagcat ctcacccgga agcgccctgg 540 ggccgaggcc accgggaagg tgaatgtcaagacgctgcgt ctgggaccgc tcagcaaggc 600 tggcttctac ctggccttcc aggaccagggtgcctgcatg gccctgctat ccctgcacct 660 cttctacaaa aagtgcgccc agctgactgtgaacctgact cgattcccgg agactgtgcc 720 tcgggagctg gttgtgcccg tggccggtagctgcgtggtg gatgccgtcc ccgcccctgg 780 ccccagcccc agcctctact gccgtgaggatggccagtgg gccgaacagc cggtcacggg 840 ctgcagctgt gctccggggt tcgaggcagctgaggggaac accaagtgcc gagcctgtgc 900 ccagggcacc ttcaagcccc tgtcaggagaagggtcctgc cagccatgcc cagccaatag 960 ccactctaac accattggat cagccgtctgccagtgccgc gtcgggtact tccgggcacg 1020 cacagacccc cggggtgcac cctgcaccacccctccttcg gctccgcgga gcgtggtttc 1080 ccgcctgaac ggctcctccc tgcacctggaatggagtgcc cccctggagt ctggtggccg 1140 agaggacctc acctacgccc tccgctgccgggagtgccga cccggaggct cctgtgcgcc 1200 ctgcggggga gacctgactt ttgaccccggcccccgggac ctggtggagc cctgggtggt 1260 ggttcgaggg ctacgtcctg acttcacctatacctttgag gtcactgcat tgaacggggt 1320 atcctcctta gccacggggc ccgtcccatttgagcctgtc aatgtcacca ctgaccgaga 1380 ggtacctcct gcagtgtctg acatccgggtgacgcggtcc tcacccagca gcttgagcct 1440 ggcctgggct gttccccggg cacccagtggggctgtgctg gactacgagg tcaaatacca 1500 tgagaagggc gccgagggtc ccagcagcgtgcggttcctg aagacgtcag aaaaccgggc 1560 agagctgcgg gggctgaagc ggggagccagctacctggtg caggtacggg cgcgctctga 1620 ggccggctac gggcccttcg gccaggaacatcacagccag acccaactgg atgagagcga 1680 gggctggcgg gagcagctgg ccctgattgcgggcacggca gtcgtgggtg tggtcctggt 1740 cctggtggtc attgtggtcg cagttctctgcctcaggaag cagagcaatg ggagagaagc 1800 agaatattcg gacaaacacg gacagtatctcatcggacat ggtactaagg tctacatcga 1860 ccccttcact tatgaagacc ctaatgaggctgtgagggaa tttgcaaaag agatcgatgt 1920 ctcctacgtc aagattgaag aggtgattggtgcaggtgag tttggcgagg tgtgccgggg 1980 gcggctcaag gccccaggga agaaggagagctgtgtggca atcaagaccc tgaagggtgg 2040 ctacacggag cggcagcggc gtgagtttctgagcgaggcc tccatcatgg gccagttcga 2100 gcaccccaat atcatccgcc tggagggcgtggtcaccaac agcatgcccg tcatgattct 2160 cacagagttc atggagaacg gcgccctggactccttcctg cggctaaacg acggacagtt 2220 cacagtcatc cagctcgtgg gcatgctgcggggcatcgcc tcgggcatgc ggtaccttgc 2280 cgagatgagc tacgtccacc gagacctggctgctcgcaac atcctagtca acagcaacct 2340 cgtctgcaaa gtgtctgact ttggcctttcccgattcctg gaggagaact cttccgatcc 2400 cacctacacg agctccctgg gaggaaagattcccatccga tggactgccc cggaggccat 2460 tgccttccgg aagttcactt ccgccagtgatgcctggagt tacgggattg tgatgtggga 2520 ggtgatgtca tttggggaga ggccgtactgggacatgagc aatcaggacg tgatcaatgc 2580 cattgaacag gactaccggc tgcccccgcccccagactgt cccacctccc tccaccagct 2640 catgctggac tgttggcaga aagaccggaatgcccggccc cgcttccccc aggtggtcag 2700 cgccctggac aagatgatcc ggaaccccgccagcctcaaa atcgtggccc gggagaatgg 2760 cggggcctca caccctctcc tggaccagcggcagcctcac tactcagctt ttggctctgt 2820 gggcgagtgg cttcgggcca tcaaaatgggaagatacgaa gaaagtttcg cagccgctgg 2880 ctttggctcc ttcgagctgg tcagccagatctctgctgag gacctgctcc gaatcggagt 2940 cactctggcg ggacaccaga agaaaatcttggccagtgtc cagcacatga agtcccaggc 3000 caagccggga accccgggtg ggacaggaggaccggccccg cagtactgac ctgcaggaac 3060 tccccacccc agggacaccg cctccccattttccggggca gagtggggac tcacagaggc 3120 ccccagccct gtgccccgct ggattgcactttgagcccgt ggggtgagga gttggcaatt 3180 tggagagaca ggatttgggg gttctgccataataggaggg gaaaatcacc ccccagccac 3240 ctcggggaac tccagaccaa gggtgagggcgcctttccct caggactggg tgtgaccaga 3300 ggaaaaggaa gtgcccaaca tctcccagcctccccaggtg cccccctcac cttgatgggt 3360 gcgttcccgc agaccaaaga gagtgtgactcccttgccag ctccagagtg ggggggctgt 3420 cccagggggc aagaaggggt gtcagggcccagtgacaaaa tcattggggt ttgtagtccc 3480 aacttgctgc tgtcaccacc aaactcaatcatttttttcc cttgtaaatg cccctccccc 3540 agctgctgcc ttcatattga aggtttttgagttttgtttt tggtcttaat ttttctcccc 3600 gttccctttt tgtttcttcg ttttgtttttctaccgtcct tgtcataact ttgtgttgga 3660 gggaacctgt ttcactatgg cctcctttgcccaagttgaa acaggggccc atcatcatgt 3720 ctgtttccag aacagtgcct tggtcatcccacatccccgg accccgcctg ggacccccaa 3780 gctgtgtcct atgaaggggt gtggggtgaggtagtgaaaa gggcggtagt tggtggtgga 3840 acccagaaac ggacgccggt gcttggaggggttcttaaat tatatttaaa aaagtaactt 3900 tttgtataaa taaaagaaaa tgggacgtgtcccagctcca ggggt 3945 5 2274 DNA Homo sapiens 5 atggcgggtc tgacggcggcggccccgcgg cccggagtcc tcctgctcct gctgtccatc 60 ctccacccct ctcggcctggaggggtccct ggggccattc ctggtggagt tcctggagga 120 gtcttttatc caggggctggtctcggagcc cttggaggag gagcgctggg gcctggaggc 180 aaacctctta agccagttcccggagggctt gcgggtgctg gccttggggc agggctcggc 240 gccttccccg cagttacctttccgggggct ctggtgcctg gtggagtggc tgacgctgct 300 gcagcctata aagctgctaaggctggcgct gggcttggtg gtgtcccagg agttggtggc 360 ttaggagtgt ctgcaggtgcggtggttcct cagcctggag ccggagtgaa gcctgggaaa 420 gtgccgggtg tggggctgccaggtgtatac ccaggtggcg tgctcccagg agctcggttc 480 cccggtgtgg gggtgctccctggagttccc actggagcag gagttaagcc caaggctcca 540 ggtgtaggtg gagcttttgctggaatccca ggagttggac cctttggggg accgcaacct 600 ggagtcccac tggggtatcccatcaaggcc cccaagctgc ctggtggcta tggactgccc 660 tacaccacag ggaaactgccctatggctat gggcccggag gagtggctgg tgcagcgggc 720 aaggctggtt acccaacagggacaggggtt ggcccccagg cagcagcagc agcggcagct 780 aaagcagcag caaagttcggtgctggagca gccggagtcc tccctggtgt tggaggggct 840 ggtgttcctg gcgtgcctggggcaattcct ggaattggag gcatcgcagg cgttgggact 900 ccagctgcag ctgcagctgcagcagcagcc gctaaggcag ccaagtatgg agctgctgca 960 ggcttagtgc ctggtgggccaggctttggc ccgggagtag ttggtgtccc aggagctggc 1020 gttccaggtg ttggtgtcccaggagctggg attccagttg tcccaggtgc tgggatccca 1080 ggtgctgcgg ttccaggggttgtgtcacca gaagcagctg ctaaggcagc tgcaaaggca 1140 gccaaatacg gggccaggcccggagtcgga gttggaggca ttcctactta cggggttgga 1200 gctgggggct ttcccggctttggtgtcgga gtcggaggta tccctggagt cgcaggtgtc 1260 cctagtgtcg gaggtgttcccggagtcgga ggtgtcccgg gagttggcat ttcccccgaa 1320 gctcaggcag cagctgccgccaaggctgcc aagtacggag tggggacccc agcagctgca 1380 gctgctaaag cagccgccaaagccgcccag tttgggttag ttcctggtgt cggcgtggct 1440 cctggagttg gcgtggctcctggtgtcggt gtggctcctg gagttggctt ggctcctgga 1500 gttggcgtgg ctcctggagttggtgtggct cctggcgttg gcgtggctcc cggcattggc 1560 cctggtggag ttgcagctgcagcaaaatcc gctgccaagg tggctgccaa agcccagctc 1620 cgagctgcag ctgggcttggtgctggcatc cctggacttg gagttggtgt cggcgtccct 1680 ggacttggag ttggtgctggtgttcctgga cttggagttg gtgctggtgt tcctggcttc 1740 ggggcaggtg cagatgagggagttaggcgg agcctgtccc ctgagctcag ggaaggagat 1800 ccctcctcct ctcagcacctccccagcacc ccctcatcac ccagggtacc tggagccctg 1860 gctgccgcta aagcagccaaatatggagca gcagtgcctg gggtccttgg agggctcggg 1920 gctctcggtg gagtaggcatcccaggcggt gtggtgggag ccggacccgc cgccgccgct 1980 gccgcagcca aagctgctgccaaagccgcc cagtttggcc tagtgggagc cgctgggctc 2040 ggaggactcg gagtcggagggcttggagtt ccaggtgttg ggggccttgg aggtatacct 2100 ccagctgcag ccgctaaagcagctaaatac ggtgctgctg gccttggagg tgtcctaggg 2160 ggtgccgggc agttcccacttggaggagtg gcagcaagac ctggcttcgg attgtctccc 2220 attttcccag gtggggcctgcctggggaaa gcttgtggcc ggaagagaaa atga 2274 6 2615 DNA Homo sapiens 6ccttttttgg cctcgacggc ggcaacccag cctccctcct aacgccctcc gcctttggga 60ccaaccaggg gagctcaagt tagtagcagc caaggagagg cgctgccttg ccaagactaa 120aaagggaggg gagaagagag gaaaaaagca agaatccccc acccctctcc cgggcggagg 180gggcgggaag agcgcgtcct ggccaagccg agtagtgtct tccactcggt gcgtctctct 240aggagccgcg cgggaaggat gctggtccgc aggggcgcgc gcgcagggcc caggatgccg 300cggggctgga ccgcgctttg cttgctgagt ttgctgcctt ctgggttcat gagtcttgac 360aacaacggta ctgctacccc agagttacct acccagggaa cattttcaaa tgtttctaca 420aatgtatcct accaagaaac tacaacacct agtacccttg gaagtaccag cctgcaccct 480gtgtctcaac atggcaatga ggccacaaca aacatcacag aaacgacagt caaattcaca 540tctacctctg tgataacctc agtttatgga aacacaaact cttctgtcca gtcacagacc 600tctgtaatca gcacagtgtt caccacccca gccaacgttt caactccaga gacaaccttg 660aagcctagcc tgtcacctgg aaatgtttca gacctttcaa ccactagcac tagccttgca 720acatctccca ctaaacccta tacatcatct tctcctatcc taagtgacat caaggcagaa 780atcaaatgtt caggcatcag agaagtgaaa ttgactcagg gcatctgcct ggagcaaaat 840aagacctcca gctgtgcgga gtttaagaag gacaggggag agggcctggc ccgagtgctg 900tgtggggagg agcaggctga tgctgatgct ggggcccagg tatgctccct gctccttgcc 960cagtctgagg tgaggcctca gtgtctactg ctggtcttgg ccaacagaac agaaatttcc 1020agcaaactcc aacttatgaa aaagcaccaa tctgacctga aaaagctggg gatcctagat 1080ttcactgagc aagatgttgc aagccaccag agctattccc aaaagaccct gattgcactg 1140gtcacctcgg gagccctgct ggctgtcttg ggcatcactg gctatttcct gatgaatcgc 1200cgcagctgga gccccacagg agaaaggctg ggcgaagacc cttattacac ggaaaacggt 1260ggaggccagg gctatagctc aggacctggg acctcccctg aggctcaggg aaaggccagt 1320gtgaaccgag gggctcagga aaacgggacc ggccaggcca cctccagaaa cggccattca 1380gcaagacaac acgtggtggc tgataccgaa ttgtgactcg gctaggtggg gcaaggctgg 1440gcagtgtccg agagagcacc cctctctgca tctgaccacg tgctaccccc atgctggagg 1500tgacatctct tacgcccaac ccttccccac tgcacacacc tcagaggctg ttcttggggc 1560cctacacctt gaggaggggc aggtaaactc ctgtccttta cacattcggc tccctggagc 1620cagactctgg tcttctttgg gtaaacgtgt gacgggggaa agccaaggtc tggagaagct 1680cccaggaaca actgatggcc ttgcagcact cacacaggac ccccttcccc taccccctcc 1740tctctgccgc aatacaggaa cccccagggg aaagatgagc ttttctaggc tacaattttc 1800tcccaggaag ctttgatttt taccgtttct tccctgtatt ttctttctct actttgagga 1860aaccaaagta accttttgca cctgctctct tgtaatgata tagccagaaa aacgtgttgc 1920cttgaaccac ttccctcatc tctcctccaa gacactgtgg acttggtcac cagctcctcc 1980cttgttctct aagttccact gagctccatg tgccccctct accatttgca gagtcctgca 2040cagttttctg gctggagcct agaacaggcc tcccaagttt taggacaaac agctcagttc 2100tagtctctct ggggccacac agaaactctt tttgggctct tttttctccc tctggatcaa 2160agtaggcagg accatgggac caggtcttgg agctgagcct ctcacctgta ctcttccgaa 2220aaatcctctt cctctgaggc tggatcctag ccttatcctc tgatctccat ggcttcctcc 2280tccctcctgc cgactcctgg gttgagctgt tgcctcagtc ccccaacaga tgcttttctg 2340tctctgcctc cctcaccctg agccccttcc ttgctctgca cccccatatg gtcatagccc 2400agatcagctc ctaaccctta tcaccagctg cctcttctgt gggtgaccca ggtccttgtt 2460tgctgttgat ttctttccag aggggttgaa cagggatcct ggtttcaatg acggttggaa 2520atagaaattt ccagagaaga gagtattggg tagatatttt ttctgaatac aaagtgatgt 2580gtttaaatac tgcaattaaa gtgatactga aacac 2615

1. A method for inducing arterial morphology in a vein, comprising:contacting endothelial cells in said vein to at least one polynucleotideencoding a gene that is capable of inducing endothelial remodeling for atime sufficient to transfer the polynucleotide into the endothelialcells.
 2. The method of claim 1, wherein said vein is a mammalian vein.3. The method of claim 1, wherein the vein is a human vein.
 4. Themethod of claim 3, wherein the vein is a saphenous vein.
 5. The methodof claim 1, wherein the gene encodes endoglin, Alk-1 or both.
 6. Themethod of claim 1, wherein the gene encodes one or more of ephrin-B2,EphB4, elastin and CD34.
 7. The method of claim 1, wherein thepolynucleotide is contained within an expression vector adapted tointroduce the polynucleotide into the cells.
 8. The method of claim 7,wherein the expression vector is a viral vector.
 9. The method of claim8, wherein the viral vector is an adenoviral vector, a herpesviralvector, a pox viral vector, or an adeno-associated viral vector.
 10. Amethod of treating a patient having an obstructed blood vessel,comprising: providing a graft comprising endothelial cells; contactingthe endothelial cells of the graft to at least one polynucleotideencoding a gene that is capable of inducing endothelial remodeling for atime sufficient to transfer the polynucleotide into the endothelialcells; removing a section of said obstructed blood vessel; and graftingthe graft in place of the removed section of said obstructed bloodvessel.
 11. The method of claim 10, wherein providing a graft comprisesharvesting a section of a vein from said patient.
 12. The method ofclaim 11, wherein the vein is a saphenous vein of said patient.
 13. Themethod of claim 10, wherein the gene encodes endoglin, Alk-1, or both.14. The method of claim 10, wherein the gene encodes one or more ofephrin-B2, EphB4, elastin and CD34.
 15. The method of claim 10, whereinthe polynucleotide is contained within an expression vector adapted tointroduce the polynucleotide into the cells.
 16. The method of claim 15,wherein the expression vector is a viral vector.
 17. The method of claim16, wherein the viral vector is an adenoviral vector, a retroviralvector, a herpesviral vector, a pox viral vector, or an adeno-associatedviral vector.
 18. A blood vessel, comprising endothelial cellscomprising an exogenously supplied polynucleotide encoding a gene thatis capable of inducing endothelial remodeling in the endothelial cells.19. A blood vessel in accordance with claim 18, wherein the vessel is asection of a mammalian vein.
 20. A blood vessel in accordance with claim19, wherein the vessel is a section of a human vein.
 21. A blood vesselin accordance with claim 20, wherein the vessel is a section of asaphenous vein.
 22. A blood vessel in accordance with claim 18, whereinthe gene encodes endoglin, Alk-1 or both.
 23. A blood vessel inaccordance with claim 18, wherein the gene encodes one or more ofephrin-B2, EphB4, elastin, and CD34.